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Biomedical subjects

Jun Wang

Publications and source records attributed to Jun Wang.

At least 109 records · Page 6Linked to original sources
PubMed2006

SiO2/Cr monolayers and formation of polyethylene films.

1,3,5-tribenzylhexahydro-1,3,5-triazine-CrCl3 (TAC-CrCl3) was supported on monodisperse Stöber silica and on commercial silica particles. The monolayers of these catalyst particles were successfully prepared with Langmuir-Blodgett technique. The effect of TAC-CrCl3 molecules on the ordering and film-forming of the silica particles was investigated. The particles arrange in domains of hexagonal ordering, but the ordering of particles in the monolayer decreases with increasing Cr loading. These hybrid monolayers are active polymerization catalysts. After polymerization, a relatively rough layer of polyethylene molecules covers the catalyst monolayer and this layer is hydrophobic. The monolayers and the polymers coating films were characterized by IR, UV-vis-NIR, and SEM. The method provides a way to fabricate polyethylene films on silica monolayers.

Journal Article↗
PubMed2006

Functional analysis of the purified anandamide-generating phospholipase D as a member of the metallo-beta-lactamase family.

In animal tissues, bioactive N-acylethanolamines including the endocannabinoid anandamide are formed from their corresponding N-acylphosphatidylethanolamines (NAPEs) by the catalysis of a specific phospholipase D (NAPE-PLD) that belongs to the metallo-beta-lactamase family. Despite its potential physiological importance, NAPE-PLD has not yet been characterized with a purified enzyme preparation. In the present study we expressed a recombinant NAPE-PLD in Escherichia coli and highly purified it. The purified enzyme was remarkably activated in a dose-dependent manner by millimolar concentrations of Mg2+ as well as Ca2+ and, hence, appeared to be constitutively active. The enzyme showed extremely high specificity for NAPEs among various glycerophospholipids but did not reveal obvious selectivity for different long chain or medium chain N-acyl species of NAPEs. These results suggested the ability of NAPE-PLD to degrade different NAPEs without damaging other membrane phospholipids. Metal analysis revealed the presence of catalytically important zinc in NAPE-PLD. In addition, site-directed mutagenesis studies were addressed to several histidine and aspartic acid residues of NAPE-PLD that are highly conserved within the metallo-beta-lactamase family. Single mutations of Asp-147, His-185, His-187, Asp-189, His-190, His-253, Asp-284, and His-321 caused abolishment or remarkable reduction of the catalytic activity. Moreover, when six cysteine residues were individually mutated to serine, only C224S showed a considerably reduced activity. The activities of L207F and H380R found as single nucleotide polymorphisms were also low. Thus, NAPE-PLD appeared to function through a mechanism similar to those of the well characterized members of this family but play a unique role in the lipid metabolism of animal tissues.

Amino Acid Sequence↗
PubMed2006

Regulation of the preprotachykinin-I gene promoter through a protein kinase A-dependent, cyclic AMP response element-binding protein-independent mechanism.

Preprotachykinin-I (PPT) gene expression is regulated by a number of stimuli that signal through cyclic AMP (cAMP)-mediated pathways. In the present study, forskolin, an adenylyl cyclase stimulator, significantly increased PPT mRNA levels in PPT-expressing RINm5F cells, an effect paralleled by an increase in PPT promoter-luciferase reporter construct activity. The forskolin-induced stimulation of PPT transcription was protein kinase A dependent (PKA), as shown by blockade with the PKA inhibitor N-[2-(p-bromocinnamylamino) ethyl]-5-isoquinolinesulfonamide. We found that the activation protein 1/cAMP response element (AP1/CRE) site centered at -196 relative to the transcription start site was important for basal and forskolin-induced PPT promoter activity. Because of the involvement of PKA and the similarity of the AP1/CRE element to consensus CRE sequences, we investigated the role of CRE-binding protein (CREB) in the regulation of the PPT promoter. Surprisingly, overexpression of a dominant-negative CREB (i.e. CREB-A) did not affect basal or forskolin-induced PPT promoter activity. Furthermore, binding of CREB to the PPT promoter AP1/CRE site was not demonstrable in electrophoretic mobility shift assays. Rather, our experiments suggested that c-Jun is a member of the complex that binds to this site. We conclude that, at least in RINm5F cells, cAMP-mediated up-regulation of PPT gene expression does not involve CREB or CREB-related transcription factor recruitment to the AP1/CRE site.

Animals↗
PubMed2006

[Comparison of wound healing after pancreaticojejunostomy with three anastomotic methods in piglets].

OBJECTIVE: To evaluate wound healing after pancreaticojejunostomy of three anastomotic methods. METHODS: Fifty-four domestic piglets were divided into three groups according to the types of anastomoses: group of end-to-end pancreaticojejunal invagination (EE group), group of binding pancreaticojejunostomy (BP group) and group of inkwell pancreaticojejunostomy (IP group). Bursting pressure, breaking strength and histopathological findings of anastomosis were assessed on operative day and on the 5th and 10th day after operation. RESULTS: Bursting pressure was (67+/-8) mm Hg, (96+/-11) mm Hg and (131+/-9) mm Hg in EE group on day 0, 5 and 10; and (140+/-8) mm Hg, (179+/-10) mm Hg and (269+/-13) mm Hg in BP group; and (102+/-10) mm Hg, (171+/-18) mm Hg and (254+/-24) mm Hg in IP group. Compare to EE group, bursting pressure of BP group and IP group were all increased with significant differences (P<0.05). Another significant difference was observed between BP group and IP group after anastomoses on operative day. Breaking strength was (4.6+/-0.6) N, (5.8+/-0.5) N and (7.1+/-0.6) N in EE group on 0 d, 5 d and 10 d; and (4.5+/-0.4) N, (6.6+/-0.4) N and (10.0+/-0.6) N in BP group; and (4.6+/-0.3) N, (6.5+/-0.4) N and (9.1+/-0.9) N in IP group. A similar value of anastomoses was shown in BP group and IP group on day 0, day 5 and day 10, but significant increase was demonstrated compared to EE group on day 5 and 10. Anastomotic site was well repaired by connective tissue and the cut surface of pancreatic stump was covered by mucosal epithelium in BP group and IP group on day 10, but the cut surface was incompletely repaired by granulation tissue and no regeneration of the epithelium was found in EE group. CONCLUSIONS: Wound healing of binding pancreaticojejunostomy and inkwell pancreaticojejunostomy is more rapid and better than end-to-end pancreaticojejunal invagination, but breaking strength of inkwell pancreaticojejunostomy is weaker than binding pancreaticojejunostomy.

Anastomosis, Surgical↗
PubMed2006

Amperometric choline biosensor fabricated through electrostatic assembly of bienzyme/polyelectrolyte hybrid layers on carbon nanotubes.

We report a flow injection amperometric choline biosensor based on the electrostatic assembly of the choline oxidase (ChO) enzyme and a bienzyme of ChO and horseradish peroxidase (HRP) onto multi-wall carbon nanotubes (MWCNT) modified glassy carbon (GC) electrodes. These choline biosensors were fabricated by immobilization of enzymes on the negatively charged MWCNT surface through alternately assembling a cationic poly(diallydimethylammonium chloride) (PDDA) layer and an enzyme layer. Using this layer-by-layer assembling approach, a bioactive nanocomposite film of PDDA/ChO/PDDA/HRP/PDDA/CNT (ChO/HRP/CNT) and PDDA/ChO/PDDA/CNT (ChO/CNT) was fabricated on the GC surface. Owing to the electrocatalytic effect of carbon nanotubes, the measurement of faradic responses resulting from enzymatic reactions has been realized at low potential with acceptable sensitivity. The ChO/HRP/CNT biosensor is more sensitive than the ChO/CNT one. Experimental parameters affecting the sensitivity of biosensors, e.g., applied potential, flow rate, etc., were optimized and potential interference was examined. The response time for this choline biosensor is fast (few seconds). The linear range of detection for the choline biosensor is from 5.0 x 10(-5) to 5.0 x 10(-3) M and the detection limit is about 1.0 x 10(-5) M.

Biosensing Techniques↗
PubMed2006

Immunosuppressive and trafficking properties of donor splenic and bone marrow dendritic cells.

BACKGROUND: Infusion of donor dendritic cells (DC) has been shown to prolong allograft survival in a number of models. However, many regimens that utilize donor DC do not consistently produced tolerance or long-term allograft survival. We hypothesized that one factor limiting the therapeutic effect of donor DC is their relative inability to traffic to recipient peripheral lymph nodes and inhibit the function of resident alloreactive T cells. METHODS: Donor strain DC isolated from the spleens or bone marrow of Flt3L-treated mice were transferred intravenously into recipients at the time of skin grafting. Where indicated, recipients were treated with an anti-CD40L antibody and CTLA4-Ig. RESULTS: Infusion of donor DC together with costimulatory blockade promoted donor-specific prolongation of skin allograft survival in mice. Perhaps due to their more immature phenotype, bone marrow DC trafficked more effectively to the spleen, bone marrow, and thymus and were associated with significantly longer allograft survival than were splenic DC. Neither population of DC trafficked well to peripheral lymph nodes. Consistent with our hypothesis, splenic but not lymph node T cells from DC-treated recipients displayed donor-specific hyporesponsiveness in vitro. CONCLUSION: These data suggest that one factor contributing to rejection following treatment with donor DC plus costimulation blockade is the persistence of donor-reactive T cells within the recipient's secondary lymphoid structures. Strategies to improve DC trafficking to these structures may enhance their therapeutic effect.

Animals↗
PubMed2006

Mitochondrial molecular clocks and the origin of the major Otocephalan clades (Pisces: Teleostei): A new insight.

The Otocephala, a clade including ostariophysan and clupeomorph teleosts, represents about a quarter of total fish species diversity, with about 1000 genera and more than 7000 species. A series of recent papers have defended that the origin of this clade and of its major groups may be significantly older than the oldest fossils of each of these groups suggest. Some of these recent papers explicitly defend a Pangean origin for some otocephalan groups such as the Siluriformes or Cypriniformes. To know whether or not the otocephalans as a whole, and particularly the mainly freshwater, cosmopolitan otophysans could have originated before the splitting of the Pangean supercontinent is of extreme importance, since otophysan fishes are among the most useful animal groups for the determination of historical continental relationships. In the present work we examined divergence times for each major otocephalan group by an analysis of complete mtDNA sequences, in order to investigate if these divergence times support the hypotheses advanced in recent studies. The complete mtDNA sequences of nine representative non-otocephalan fish species and of twenty-one representative otocephalan species was compared. The present study is thus, among the studies dealing with molecular divergence times of teleosts, the one in which a greater number of otocephalan species are included. The divergence times obtained support that the major otocephalan groups had a much older origin than the oldest fossil records available for these groups suggest. The origin of the Otocephala is estimated as having occurred about 282 Mya, with the origin of the Otophysi being estimated at about 251 Mya.

Animals↗
PubMed2006

High-throughput screening for the asymmetric transformation reaction of L-histidine to D-histidine by capillary array electrophoresis.

Asymmetric transformation reaction of L-histidine to D-histidine was studied by homemade capillary array electrophoresis for the first time. The enantiomeric excess value of asymmetric histidine products can be directly determined from the electrophoretogram of capillary array electrophoresis. The experiment results showed that the optimized asymmetric transformation reaction condition was in the presence of salicylaldehyde as catalyst and acetic acid as solvent.

Aldehydes↗
PubMed2006

J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase from Bacillus laterosporus J1, is a member of the alpha/beta-hydrolase fold superfamily.

J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase (GCA) isolated from Bacillus laterosporus J1, has been conventionally grouped as the only member of class V GCA, although its amino acid sequence shares less than 10% identity with members of other classes of GCA. Instead, it shows higher sequence similarities with Rhodococcus sp. strain MB1 cocaine esterase (RhCocE) and Acetobacter turbidans alpha-amino acid ester hydrolase (AtAEH), members of the alpha/beta-hydrolase fold superfamily. Homology modeling and secondary structure prediction indicate that the N-terminal region of J1 acylase has an alpha/beta-hydrolase folding pattern. The catalytic triads in RhCocE and AtAEH were identified in J1 acylase as S125, D264 and H309. Mutations to alanine at these positions were found to completely inactivate the enzyme. These results suggest that J1 acylase is a member of the alpha/beta-hydrolase fold superfamily with a serine-histidine-aspartate catalytic triad.

Amidohydrolases↗
PubMed2006

Single-channel properties of volume-sensitive Cl- channel in ClC-3-deficient cardiomyocytes.

It is controversial whether the ClC-3 protein, which is one of the voltage-dependent chloride channel ClC family members, is a candidate for the volume-sensitive outwardly rectifying (VSOR) Cl(-) channel per se or its regulator. Here, for the first time, we examined the single-channel properties of the VSOR Cl(-) channel in ventricular myocytes isolated from ClC-3-deficient mice. The single-channel current induced by cell swelling exhibited Cl(-) selectivity, mild outward rectification, and an intermediate unitary conductance (around 38 pS). A Cl(-) channel blocker, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), reversibly inhibited the outward current. These single-channel properties were identical with those in ClC-3 expressing wild-type ventricular myocytes. These results indicate that the single-channel activity of the VSOR Cl(-) channel is independent of the expression of ClC-3 proteins in mouse ventricular myocytes.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2006

Splice variants of the NR1 subunit differentially induce NMDA receptor-dependent gene expression.

Subunits of the NMDA receptor (NMDAR) associate with many postsynaptic proteins that substantially broaden its signaling capacity. Although much work has been focused on the signaling of NR2 subunits, little is known about the role of the NR1 subunit. We set out to elucidate the role of the C terminus of the NR1 subunit in NMDAR signaling. By introducing a C-terminal deletion mutant of the NR1 subunit into cultured neurons from NR1(-/-) mice, we found that the C terminus was essential for NMDAR inactivation, downstream signaling, and gene expression, but not for global increases in intracellular Ca2+. Therefore, whereas NMDARs can increase Ca2+ throughout the neuron, NMDAR-dependent signaling, both local and long range, requires coupling through the NR1 C terminus. Two major NR1 splice variants differ by the presence or absence of a C-terminal domain, C1, which is determined by alternative splicing of exon 21. Analysis of these two variants showed that removal of this domain significantly reduced the efficacy of NMDAR-induced gene expression without affecting receptor inactivation. Thus, the NR1 C terminus couples to multiple downstream signaling pathways that can be modulated selectively by RNA splicing.

Alternative Splicing↗
PubMed2006

Phage display selection of peptides that inhibit metastasis ability of gastric cancer cells with high liver-metastatic potential.

Organ-specific metastasis is an important character of cancer cells. Cancer cells that can metastasize to a special organ were thought to have different proteins in cell membrane, which might have potential utility as diagnostic markers and therapeutic targets. In the present work, based on high liver-metastatic gastric cancer cells, XGC9811-L, a screening approach with phage displayed peptide library, was successfully used to isolate 8-mer peptide ligands binding to the target cells. The phage20 had the highest binding efficiency to XGC9811-L cells, which also displayed remarkable cell specificity. Peptide20 that was displayed on phage20 could suppress the motility and invasion of XGC9811-L significantly. The adhesive ability of XGC9811-L to collagen IV was also inhibited by peptide20. Furthermore, phage20 could significantly reduce the incidence of liver metastasis of gastric cancer transplanted into nude mice and was also beneficial for the reduction the number of metastatic nodules in the liver. In conclusion, the phage display is an effective method to screen for the new molecules associated with organ-specific metastasis. The selected peptide20 can reverse the liver metastasis behavior of the gastric cancer cells.

Amino Acid Sequence↗
PubMed2006

Cleavage targets and the D-arginine-based inhibitors of the West Nile virus NS3 processing proteinase.

Mosquito-borne WNV (West Nile virus) is an emerging global threat. The NS3 proteinase, which is essential for the proteolytic processing of the viral polyprotein precursor, is a promising drug target. We have isolated and biochemically characterized the recombinant, highly active NS3 proteinase. We have determined that the NS3 proteinase functions in a manner that is distantly similar to furin in cleaving the peptide and protein substrates. We determined that aprotinin and D-arginine-based 9-12-mer peptides are potent inhibitors of WNV NS3 with K(i) values of 26 nM and 1 nM respectively. Consistent with the essential role of NS3 activity in the life cycle of WNV and with the sensitivity of NS3 activity to the D-arginine-based peptides, we showed that nona-D-Arg-NH2 reduced WNV infection in primary neurons. We have also shown that myelin basic protein, a deficiency of which is linked to neurological abnormalities of the brain, is sensitive to NS3 proteolysis in vitro and therefore this protein represents a convenient test substrate for the studies of NS3. A three-dimensional model of WNV NS3 that we created may provide a structural guidance and a rationale for the subsequent design of fine-tuned inhibitors. Overall, our findings represent a foundation for in-depth mechanistic and structural studies as well as for the design of novel and efficient inhibitors of WNV NS3.

Amino Acid Sequence↗
PubMed2006

Sonocatalytic degradation of methyl parathion in the presence of nanometer and ordinary anatase titanium dioxide catalysts and comparison of their sonocatalytic abilities.

The degradation of methyl parathion (O,O-dimethyl-O-(4-nitrophenyl)-phosphorothioate) using anatase titanium dioxide (TiO(2)) powder as heterogeneous sonocatalysts is reported. The influences of reaction parameters such as the species of TiO(2) sonocatalysts, methyl parathion concentrations, TiO(2) adding amount, pH, ultrasonic intensity, ultrasonic frequency and temperature have been investigated and the optimal conditions for eliminating methyl parathion have been identified. The efficiencies of sonocatalytic degradation in both nanometer and ordinary anatase systems are compared and the results indicate that the sonocatalytic activity of nanometer anatase TiO(2) powder is better than that of ordinary anatase TiO(2) powder. The primary degradation and the total mineralization of methyl parathion have been monitored by high performance liquid chromatography (HPLC) and UV-vis spectra, respectively. Methyl parathion got destroyed to some extent in both nanometer and ordinary anatase systems under ultrasonic irradiation. The kinetics for the degradation process of methyl parathion follows the first-order reaction. The degradation ratio of methyl parathion surpassed 90% within 50min in the optimal experiment conditions.

Catalysis↗
PubMed2006

TreeFam: a curated database of phylogenetic trees of animal gene families.

TreeFam is a database of phylogenetic trees of gene families found in animals. It aims to develop a curated resource that presents the accurate evolutionary history of all animal gene families, as well as reliable ortholog and paralog assignments. Curated families are being added progressively, based on seed alignments and trees in a similar fashion to Pfam. Release 1.1 of TreeFam contains curated trees for 690 families and automatically generated trees for another 11 646 families. These represent over 128 000 genes from nine fully sequenced animal genomes and over 45 000 other animal proteins from UniProt; approximately 40-85% of proteins encoded in the fully sequenced animal genomes are included in TreeFam. TreeFam is freely available at http://www.treefam.org and http://treefam.genomics.org.cn.

Animals↗